Georgia State University ScholarWorks @ Georgia State University Chemistry Dissertations Department of Chemistry 12-14-2017 New Chemical Strategies for prodrug preparations: from sulfide ot doxorubicin Yueqin Zheng Georgia State University Follow this and additional works at: https://scholarworks.gsu.edu/chemistry_diss Recommended Citation Zheng, Yueqin, "New Chemical Strategies for prodrug preparations: from sulfide ot doxorubicin." Dissertation, Georgia State University, 2017. https://scholarworks.gsu.edu/chemistry_diss/139 This Dissertation is brought to you for free and open access by the Department of Chemistry at ScholarWorks @ Georgia State University. It has been accepted for inclusion in Chemistry Dissertations by an authorized administrator of ScholarWorks @ Georgia State University. For more information, please contact [email protected]. NEW CHEMICAL STRATEGIES FOR PRODRUG PREPARATIONS: FROM SULFIDE TO DOXORUBICIN by YUEQIN ZHENG Under the Direction of Binghe Wang, PhD ABSTRACT Prodrug is an often-used approach to facilitate the delivery of an active drug to an appropriate site with targeted release whenever possible. Proper prodrug design often relies on a few essential requirements: prodrug stability, triggered release, and selectivity. In the last few decades, there have been impressive progress in prodrug development. However, the delivery of gasotransmitters and ensuring linker stability while allowing drug release at the desired site of action are among remaining challenges. The dissertation work focuses on developing new chemical strategies to address these two issues using hydrogen sulfide (H2S) as a model for gasotransmitters and doxorubicin as a model for anticancer compounds. In the gasotransmiter part of the project, we developed a “trimethyl lock”-lactonization based approach to deliver pure H2S, an esterase-sensitive acetal approach to deliver persulfide, and an enrichment-triggered release method to deliver doxorubicin. The therapeutic effects of these prodrugs were evaluated in a combination of in vitro and in vivo models. The concepts described should be generally applicable and should be very useful to those interested in prodrug design. INDEX WORDS: Hydrogen sulfide, Persulfide prodrugs, Prodrug activation, Drug delivery, Click and release, Enrichment to release, Kinetic control drug release. NEW CHEMICAL STRATEGIES FOR PRODRUG PREPARATIONS: FROM SULFIDE TO DOXORUBICIN by YUEQIN ZHENG A Dissertation Submitted in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy in the College of Arts and Sciences Georgia State University 2017 Copyright by Yueqin Zheng 2017 NEW CHEMICAL STRATEGIES FOR PRODRUG PREPARATIONS: FROM SULFIDE TO DOXORUBICIN by YUEQIN ZHENG Committee Chair: Binghe Wang Committee: Jun Yin Donald Hamelberg Electronic Version Approved: Office of Graduate Studies College of Arts and Sciences Georgia State University December 2017 iv DEDICATION I would like to dedicate this dissertation to my parents (Lisheng Zheng and Zhaolan Zeng) and my wife (Weiwei Guo). Without you, I would not be where I am today. You gave me spiritual as well as material support in my pursuit of knowledge. Your love, kindness, encouragement and care cannot be compared by anything in the world. I also dedicate my work to my daughter (Liya Zheng). Welcome to this world and you give me endless happiness and infinite power. 1 ACKNOWLEDGEMENTS I owe my deepest gratitude to my advisor, Dr. Binghe Wang, whose encouragement, guidance and support enable me to be an independent chemist and scientist. His personality and ways of doing things not only influence my research but most importantly teach me how to be a successful person. I would also like to thank Dr. Donald Hamelberg, Dr. Jun Yin, Dr. Alfons Leopold Baumstark, Dr. Stuart Anthony Allison, Dr. Hao Xu, and Dr. Ning Fang. Their advices guide me throughout my PhD pursuing time. I want to show my great gratitude to all other faculty members and staffs for their help. I am indebted to many of my colleges for their sensational supports and fruitful advices. Especially, I am grateful to Dr. Weixuan Chen and Dr. Chaofeng Dai for their mentoring and assistance in many ways, to Bingchen Yu, Zhixiang Pan, Manjusha Roy Choudhury, Zhengnan Yuan, Vayou Chittavong for their generous help, to Dr. Kaili Ji for all biological work, to Dr. Siming Wang and Lifang Wang for mass spectrometry work, to Mengyuann Zhu for computational modeling, and to Dr. Jianmei Cui, Dr. Danzhu Wang, Dr. Bowen Ke, Dr. Yunfeng Cheng, Dr. Xingyue Ji, Dr. Krishna Damera, Dr. Hanjing Peng, Dr. Sarah Burroughs, Dr. Alexander Draganov, Dr. Ke Wang, Dr. Jalisa Nicole Holmes, Dr. Xiaoxiao Yang, Abiodun Anifowose, Jun Zhang, Ladie Kimberly Caya De La Cruz, Wenyi Wang and all other group members for their discussions and helps in research. I am also grateful for the financial support from the National Institutes of Health and the Department of Chemistry at Georgia State University. 2 TABLE OF CONTENTS ACKNOWLEDGEMENTS ......................................................................................................... 1 LIST OF TABLES ........................................................................................................................ 5 LIST OF FIGURES ...................................................................................................................... 6 LIST OF SCHEMES .................................................................................................................... 8 1 DEVELOPMENT OF ESTERASE-SENSITIVE PRODRUGS WITH TUNABLE RELEASE RATES AND DIRECT GENERATION OF HYDROGEN SULFIDE ... 9 1.1 INTRODUCTION ........................................................................................................ 9 1.1.1 The homeostasis of hydrogen sulfide and its pharmacological effect ....... 10 1.1.2 Current Challenges. ..................................................................................... 15 1.1.3 Current H2S donors ..................................................................................... 21 1.2 Results and Discussions ............................................................................................. 24 1.3 Conclusion ................................................................................................................... 32 1.4 Experimental part ...................................................................................................... 32 1.4.1 Synthesis of HPs........................................................................................... 33 1.4.2 H2S release from HPs .................................................................................. 49 1.4.3 Kinetic studies of esterase trigger lactone formation ................................. 51 1.4.4 Stability Studies of the thioacid group ........................................................ 53 1.4.5 Cell culture ................................................................................................... 55 1.5 Acknowledgement ...................................................................................................... 57 3 2 DEVEPMENT OF A NOVEL ESTERASE-SENSITIVE PRODRUG APPROACH FOR CONTROLLABLE DELIVERY OF PERSULFIDE SPECIES ....................... 58 2.1 Introduction ................................................................................................................ 58 2.2 Results and Discussions ............................................................................................. 60 2.3 Conclusions ................................................................................................................. 70 2.4 Experimental part ...................................................................................................... 70 2.1.1 Synthesis ....................................................................................................... 71 2.4.2 Kinetic studies of P1-6 (Ps) in the presence of esterase. ............................ 79 2.4.3 Mechanism study of H2S release from “acetal” based H2S prodrugs using HPLC. ....................................................................................................................... 82 2.4.4 Cell culture ................................................................................................... 83 2.4.5 LC-MS studies of persulfide reactivity with S-methyl methanethiosulfonate (MMTS). ....................................................................................................................... 87 2.4.7 Myocardial Ischemia-Reperfusion Protocol ............................................... 91 2.4.8 Sulfane Sulfur Measurement Protocol ....................................................... 93 2.5 Acknowledgement ...................................................................................................... 94 3 AN ENRICHMENT-TRIGGERED PRODRUG ACTIVATION STRATEGY DEMONSTRATED USING A CLICK, CYCLIZE, AND RELEASE SYSTEM FOR DELIVERY OF DOXORUBICIN IN MITOCHONDRIA ............................... 95 3.1 Introduction ................................................................................................................ 95 3.2 Results and Discussions ............................................................................................. 97 3.3 Conclusion ................................................................................................................. 106 4 3.4 Experiment part ......................................................................................................